pNV-sgRNA-Cas9-2A-GFP

                            LOCUS       dna                      9479 bp                                   
FEATURES             Location/Qualifiers
     Reporter        239..398
                     /gene="lacZ_a reporter"
     misc_binding    396..401
                     /dbxref="REBASE:EcoRI"
     misc_binding    408..413
                     /dbxref="REBASE:KpnI"
     misc_binding    414..419
                     /dbxref="REBASE:NruI"
     misc_binding    425..430
                     /dbxref="REBASE:XbaI"
     ORF             426..1358
                     /sequence="ORF_2 rf(4)"
     Promoter        441..1618
                     /gene="EF1a prom"
     misc_binding    1405..1410
                     /dbxref="REBASE:XhoI"
     ORF             1430..4195
                     /sequence="ORF_3 rf(5)"
     ORF             1628..6613
                     /sequence="ORF_1 rf(2)"
     Reporter        5894..6610
                     /gene="EGFP reporter"
     misc_binding    7204..7209
                     /dbxref="REBASE:BamHI"
     misc_binding    7208..7213
                     /dbxref="REBASE:SmaI"
     misc_binding    7208..7213
                     /dbxref="REBASE:XmaI"
     misc_binding    7217..7222
                     /dbxref="REBASE:AccI"
     misc_binding    7217..7222
                     /dbxref="REBASE:HincII"
     misc_binding    7240..7245
                     /dbxref="REBASE:HindIII"
     Promoter        7307..7336
                     /gene="lac prom"
     Rep_Origin      7645..8264
                     /gene="pBR322 origin"
     Marker          8419..9279
                     /gene="amp marker"
     ORF             8419..9279
                     /sequence="ORF_4 rf(6)"
     Promoter        9321..9349
                     /gene="amp prom"

        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca     
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg     
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc     
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc     
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat     
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt     
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt acctcgcgaa     
      421 tgcatctaga tatccctagc ggctccggtg cccgtcagtg ggcagagcgc acatcgccca     
      481 cagtccccga gaagttgggg ggaggggtcg gcaattgaac cggtgcctag agaaggtggc     
      541 gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc gagggtgggg     
      601 gagaaccgta tataagtgca gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg     
      661 ccagaacaca ggtaagtgcc gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg     
      721 gcccttgcgt gccttgaatt acttccactg gctgcagtac gtgattcttg atcccgagct     
      781 tcgggttgga agtgggtggg agagttcgag gccttgcgct taaggagccc cttcgcctcg     
      841 tgcttgagtt gaggcctggc ctgggcgctg gggccgccgc gtgcgaatct ggtggcacct     
      901 tcgcgcctgt ctcgctgctt tcgataagtc tctagccatt taaaattttt gatgacctgc     
      961 tgcgacgctt tttttctggc aagatagtct tgtaaatgcg ggccaagatc tgcacactgg     
     1021 tatttcggtt tttggggccg cgggcggcga cggggcccgt gcgtcccagc gcacatgttc     
     1081 ggcgaggcgg ggcctgcgag cgcggccacc gagaatcgga cgggggtagt ctcaagctgg     
     1141 ccggcctgct ctggtgcctg gcctcgcgcc gccgtgtatc gccccgccct gggcggcaag     
     1201 gctggcccgg tcggcaccag ttgcgtgagc ggaaagatgg ccgcttcccg gccctgctgc     
     1261 agggagctca aaatggagga cgcggcgctc gggagagcgg gcgggtgagt cacccacaca     
     1321 aaggaaaagg gcctttccgt cctcagccgt cgcttcatgt gactccacgg agtaccgggc     
     1381 gccgtccagg cacctcgatt agttctcgag cttttggagt acgtcgtctt taggttgggg     
     1441 ggaggggttt tatgcgatgg agtttcccca cactgagtgg gtggagactg aagttaggcc     
     1501 agcttggcac ttgatgtaat tctccttgga atttgccctt tttgagtttg gatcttggtt     
     1561 cattctcaag cctcagacag tggttcaaag tttttttctt ccatttcagg tgtcgtgagc     
     1621 gatcgccatg gccccaaaga agaagcggaa ggtcggtatc cacggagtcc cagcagccga     
     1681 caagaagtac agcatcggcc tggacatcgg caccaactct gtgggctggg ccgtgatcac     
     1741 cgacgagtac aaggtgccca gcaagaaatt caaggtgctg ggcaacaccg accggcacag     
     1801 catcaagaag aacctgatcg gagccctgct gttcgacagc ggcgaaacag ccgaggccac     
     1861 ccggctgaag agaaccgcca gaagaagata caccagacgg aagaaccgga tctgctatct     
     1921 gcaagagatc ttcagcaacg agatggccaa ggtggacgac agcttcttcc acagactgga     
     1981 agagtccttc ctggtggaag aggataagaa gcacgagcgg caccccatct tcggcaacat     
     2041 cgtggacgag gtggcctacc acgagaagta ccccaccatc taccacctga gaaagaaact     
     2101 ggtggacagc accgacaagg ccgacctgcg gctgatctat ctggccctgg cccacatgat     
     2161 caagttccgg ggccacttcc tgatcgaggg cgacctgaac cccgacaaca gcgacgtgga     
     2221 caagctgttc atccagctgg tgcagaccta caaccagctg ttcgaggaaa accccatcaa     
     2281 cgccagcggc gtggacgcca aggccatcct gtctgccaga ctgagcaaga gcagacggct     
     2341 ggaaaatctg atcgcccagc tgcccggcga gaagaagaat ggcctgttcg gcaacctgat     
     2401 tgccctgagc ctgggcctga cccccaactt caagagcaac ttcgacctgg ccgaggatgc     
     2461 caaactgcag ctgagcaagg acacctacga cgacgacctg gacaacctgc tggcccagat     
     2521 cggcgaccag tacgccgacc tgtttctggc cgccaagaac ctgtccgacg ccatcctgct     
     2581 gagcgacatc ctgagagtga acaccgagat caccaaggcc cccctgagcg cctctatgat     
     2641 caagagatac gacgagcacc accaggacct gaccctgctg aaagctctcg tgcggcagca     
     2701 gctgcctgag aagtacaaag agattttctt cgaccagagc aagaacggct acgccggcta     
     2761 cattgacggc ggagccagcc aggaagagtt ctacaagttc atcaagccca tcctggaaaa     
     2821 gatggacggc accgaggaac tgctcgtgaa gctgaacaga gaggacctgc tgcggaagca     
     2881 gcggaccttc gacaacggca gcatccccca ccagatccac ctgggagagc tgcacgccat     
     2941 tctgcggcgg caggaagatt tttacccatt cctgaaggac aaccgggaaa agatcgagaa     
     3001 gatcctgacc ttccgcatcc cctactacgt gggccctctg gccaggggaa acagcagatt     
     3061 cgcctggatg accagaaaga gcgaggaaac catcaccccc tggaacttcg aggaagtggt     
     3121 ggacaagggc gcttccgccc agagcttcat cgagcggatg accaacttcg ataagaacct     
     3181 gcccaacgag aaggtgctgc ccaagcacag cctgctgtac gagtacttca ccgtgtataa     
     3241 cgagctgacc aaagtgaaat acgtgaccga gggaatgaga aagcccgcct tcctgagcgg     
     3301 cgagcagaaa aaggccatcg tggacctgct gttcaagacc aaccggaaag tgaccgtgaa     
     3361 gcagctgaaa gaggactact tcaagaaaat cgagtgcttc gactccgtgg aaatctccgg     
     3421 cgtggaagat cggttcaacg cctccctggg cacataccac gatctgctga aaattatcaa     
     3481 ggacaaggac ttcctggaca atgaggaaaa cgaggacatt ctggaagata tcgtgctgac     
     3541 cctgacactg tttgaggaca gagagatgat cgaggaacgg ctgaaaacct atgcccacct     
     3601 gttcgacgac aaagtgatga agcagctgaa gcggcggaga tacaccggct ggggcaggct     
     3661 gagccggaag ctgatcaacg gcatccggga caagcagtcc ggcaagacaa tcctggattt     
     3721 cctgaagtcc gacggcttcg ccaacagaaa cttcatgcag ctgatccacg acgacagcct     
     3781 gacctttaaa gaggacatcc agaaagccca ggtgtccggc cagggcgata gcctgcacga     
     3841 gcacattgcc aatctggccg gcagccccgc cattaagaag ggcatcctgc agacagtgaa     
     3901 ggtggtggac gagctcgtga aagtgatggg ccggcacaag cccgagaaca tcgtgatcga     
     3961 aatggccaga gagaaccaga ccacccagaa gggacagaag aacagccgcg agagaatgaa     
     4021 gcggatcgaa gagggcatca aagagctggg cagccagatc ctgaaagaac accccgtgga     
     4081 aaacacccag ctgcagaacg agaagctgta cctgtactac ctgcagaatg ggcgggatat     
     4141 gtacgtggac caggaactgg acatcaaccg gctgtccgac tacgatgtgg accatatcgt     
     4201 gcctcagagc tttctgaagg acgactccat cgacaacaag gtgctgacca gaagcgacaa     
     4261 gaaccggggc aagagcgaca acgtgccctc cgaagaggtc gtgaagaaga tgaagaacta     
     4321 ctggcggcag ctgctgaacg ccaagctgat tacccagaga aagttcgaca atctgaccaa     
     4381 ggccgagaga ggcggcctga gcgaactgga taaggccggc ttcatcaaga gacagctggt     
     4441 ggaaacccgg cagatcacaa agcacgtggc acagatcctg gactcccgga tgaacactaa     
     4501 gtacgacgag aatgacaagc tgatccggga agtgaaagtg atcaccctga agtccaagct     
     4561 ggtgtccgat ttccggaagg atttccagtt ttacaaagtg cgcgagatca acaactacca     
     4621 ccacgcccac gacgcctacc tgaacgccgt cgtgggaacc gccctgatca aaaagtaccc     
     4681 taagctggaa agcgagttcg tgtacggcga ctacaaggtg tacgacgtgc ggaagatgat     
     4741 cgccaagagc gagcaggaaa tcggcaaggc taccgccaag tacttcttct acagcaacat     
     4801 catgaacttt ttcaagaccg agattaccct ggccaacggc gagatccgga agcggcctct     
     4861 gatcgagaca aacggcgaaa ccggggagat cgtgtgggat aagggccggg attttgccac     
     4921 cgtgcggaaa gtgctgagca tgccccaagt gaatatcgtg aaaaagaccg aggtgcagac     
     4981 aggcggcttc agcaaagagt ctatcctgcc caagaggaac agcgataagc tgatcgccag     
     5041 aaagaaggac tgggacccta agaagtacgg cggcttcgac agccccaccg tggcctattc     
     5101 tgtgctggtg gtggccaaag tggaaaaggg caagtccaag aaactgaaga gtgtgaaaga     
     5161 gctgctgggg atcaccatca tggaaagaag cagcttcgag aagaatccca tcgactttct     
     5221 ggaagccaag ggctacaaag aagtgaaaaa ggacctgatc atcaagctgc ctaagtactc     
     5281 cctgttcgag ctggaaaacg gccggaagag aatgctggcc tctgccggcg aactgcagaa     
     5341 gggaaacgaa ctggccctgc cctccaaata tgtgaacttc ctgtacctgg ccagccacta     
     5401 tgagaagctg aagggctccc ccgaggataa tgagcagaaa cagctgtttg tggaacagca     
     5461 caagcactac ctggacgaga tcatcgagca gatcagcgag ttctccaaga gagtgatcct     
     5521 ggccgacgct aatctggaca aagtgctgtc cgcctacaac aagcaccggg ataagcccat     
     5581 cagagagcag gccgagaata tcatccacct gtttaccctg accaatctgg gagcccctgc     
     5641 cgccttcaag tactttgaca ccaccatcga ccggaagagg tacaccagca ccaaagaggt     
     5701 gctggacgcc accctgatcc accagagcat caccggcctg tacgagacac ggatcgacct     
     5761 gtctcagctg ggaggcgaca aaaggccggc ggccacgaaa aaggccggcc aggcaaaaaa     
     5821 gaaaaagacg cgtcagtgta ctaattatgc tctcttgaaa ttggctggag atgttgagag     
     5881 caacccaggt cccatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct     
     5941 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg     
     6001 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt     
     6061 gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc     
     6121 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga     
     6181 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga     
     6241 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa     
     6301 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga     
     6361 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag     
     6421 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct     
     6481 gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg     
     6541 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga     
     6601 gctgtacaag taagtttaaa cgtcgagctg tgccttctag ttgccagcca tctgttgttt     
     6661 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat     
     6721 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg     
     6781 tggggcagga cagcaagggg gaggattggg aagagaatag caggcatgct ggggacaatt     
     6841 ggagggccta tttcccatga ttccttcata tttgcatata cgatacaagg ctgttagaga     
     6901 gataattaga attaatttga ctgtaaacac aaagatatta gtacaaaata cgtgacgtag     
     6961 aaagtaataa tttcttgggt agtttgcagt tttaaaatta tgttttaaaa tggactatca     
     7021 tatgcttacc gtaacttgaa agtatttcga tttcttgggt ttatatatct tgtggaaagg     
     7081 acgaaacacc ggagtcttca ccggtgaaga cgcgttttag agctagaaat agcaagttaa     
     7141 aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct tttttttgat     
     7201 atcggatccc gggcccgtcg actgcagagg cctgcatgca agcttggcgt aatcatggtc     
     7261 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg     
     7321 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt     
     7381 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg     
     7441 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga     
     7501 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat     
     7561 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca     
     7621 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc     
     7681 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata     
     7741 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc     
     7801 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc     
     7861 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga     
     7921 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc     
     7981 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag     
     8041 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag     
     8101 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag     
     8161 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca     
     8221 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga     
     8281 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat     
     8341 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga     
     8401 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg     
     8461 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga     
     8521 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc     
     8581 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac     
     8641 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc     
     8701 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc     
     8761 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc     
     8821 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt     
     8881 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc     
     8941 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg     
     9001 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag     
     9061 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat     
     9121 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc     
     9181 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa     
     9241 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta     
     9301 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa     
     9361 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga     
     9421 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtc